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The study was performed on six male chinchillas. The animals were anaesthetised with ether and the anaesthesia was deepened with nembuthal injected intraperitoneally. The chinchillas were then transcardially perfused with 0.4 L of 4% buffered paraformaldehyde, and testes, epididymides, and vasa deferentia were collected. The tunica albugínea from one testis from each chinchilla was stained as whole-mount preparation. The tissues were cut into 12 µm-thick cryostat sections, and processed for double-immunofluorescence method. In all organs studied, the most abundant nerve fibres were dopamine ß hydroxylase positive (DßH⁺). Some of them contained neuropeptide Y (NPY). Sporadically NPY-positive-only nerve fibres were found. Single DßH⁺ nerve terminals contained also galanine. Small numbers of the nerve fibres supplying studied organs were stained for substance P (SP) and calitonin gene related peptide (CGRP). Almost all SP⁺ fibres were also CGRP⁺, whereas single CGRP⁺ nerves were SP- immunonegative. Some nerve terminals were immunoreactive to vesicular acetylcholine transporter and vasoactive intestinal peptide. The organs studied were innervated unevenly. The highest density of the nerves was found in the areas of the tunica albuginea adjacent to the mesorchial border of the testis and their number gradually decreased towards the free border of the gonad. None of the vascular tissue of the testicular parenchyma was free of the nerve fibres, except sporadically encountered DßH⁺ nerves which supply seminiferous tubules. Within the head of the epididymis a moderate number of nerve terminals were found, but in the body and tail of the organ the number of nerves gradually increased. The vas deferens was supplied with very numerous nerve fibres. There were no differences in the density of the innervation between the funicular and abdominal part of the vas deferens.
This study describes the expression of DßH as a marker of sympathetic (catecholaminergic) innervation in the vestigial uterus of adult European bison (Eb) bulls. Cryo-sectioned uteri were examined by fluorescent immunohistochemistry with the use of primary rabbit DßH polyclonal antibodies. The DßH-immunocomplexes (antigens/polyclonals) were visualised by secondary donkey antirabbit biotinylated IγG, and then with cyanine (CY™3)-conjugated streptavidin. The DßH immunodetection with CY™3 for the Eb uterine sections was performed in parallel to a positive control (porcine sympathetic paravertebral ganglion), and a negative control (without primary anti-DßH polyclonals). This is the first paper describing the identification of DßH expression within the vestigial uterus of the adult Eb bulls. The distribution pattern of DßH expression localised within immunoreactive (IR) uterine nerve fibres of the bulls resembled that in the control female uterus. The DßH-IR nerve fibres were identified in the entire cross-section of each uterus, with generally higher density in the myometrium than in the endometrium. The ratio of the endometrial/myometrial DßH-expression (percentage of area with IR-signals) was comparable in all males and females (0.78 and 0.64, respectively). However, the DßH-expression area (%) was significantly lower within the male uterine endometrial (P<0.01) and myometrial regions (P<0.001) comparing to the female counterparts. Presumably, the DßH-IR expression within the male Eb uterus is associated with a trophic effect of noradrenaline released by sympathetic nerve fibres influencing nutrient supply of this vestigial organ in the Eb bulls.
The uterine cervix-projecting neurons located in the lumbar paravertebral ganglia were identified by retrograde tracing. These contained immunoreactivity to TH and DBH. No immunoreactivity to GAL, VIP and SP was found in the neurons. Extirpation of the uterus reduced the expression of TH and induced the expression of GAL in the neurons. Expression of other substances studied was unchanged.
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